| A method for the determination of enzyme mass loading on an electrode surface through radioisotope labelling. | |
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MedLine Citation:
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PMID: 12392945 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A direct method has been developed for the quantitation of the amount of immobilised enzymes on biosensor surfaces. This quantity is of key importance in establishing the activity, kinetics and optimal immobilisation conditions in the construction of both amperometric and optical biosensors. Recombinant L-lactate dehydrogenase incorporating both a biosynthetically introduced radiolabel, 3H-leucine, and a hexahistidine peptide tag was immobilised on a poly(aniline) composite film and then quantitated by liquid scintillation counting. It was found that enzyme mass loading was proportional to the concentration of LDH in solution, and also depended on the morphology of the composite film. The LDH mass loading on the composite film doubled when a surface cysteine containing variant was used, possibly due to the covalent attachment of the cysteine to the diiminoquinoid rings of the poly(aniline). |
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Authors:
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Catherine M Halliwell; Evelyne Simon; Chee-Seng Toh; Philip N Bartlett; Anthony E G Cass |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Biosensors & bioelectronics Volume: 17 ISSN: 0956-5663 ISO Abbreviation: Biosens Bioelectron Publication Date: 2002 Dec |
Date Detail:
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Created Date: 2002-10-23 Completed Date: 2003-06-18 Revised Date: 2009-07-14 |
Medline Journal Info:
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Nlm Unique ID: 9001289 Medline TA: Biosens Bioelectron Country: England |
Other Details:
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Languages: eng Pagination: 965-72 Citation Subset: IM |
Affiliation:
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Department of Biological Sciences, Imperial College of Science, Technology and Medicine, Exhibition Road, London SW7 2AY, UK. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Aniline Compounds* Biosensing Techniques Catalysis Coated Materials, Biocompatible / chemistry Electrochemistry / methods* Enzyme Activation Enzymes, Immobilized / analysis*, chemistry Equipment Design / methods* Isotope Labeling / instrumentation, methods* L-Lactate Dehydrogenase / analysis*, chemistry Leucine / chemistry Microscopy, Electron, Scanning Recombinant Proteins / analysis, chemistry Scintillation Counting / methods Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
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0/Aniline Compounds; 0/Coated Materials, Biocompatible; 0/Enzymes, Immobilized; 0/Recombinant Proteins; 0/polyaniline; 61-90-5/Leucine; EC 1.1.1.27/L-Lactate Dehydrogenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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